The EC cells in logarithmic growth phase were collected for transfection with miR-93-5p mimic, miR-93-5p inhibitor and miR-93-5p mimic + oe-TGFβR2 as well as their corresponding controls (obtained from Guangzhou RiboBio Co., Ltd.) using Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.), respectively. The transfection concentration was 50 nM. Following 6 h of incubation with 5% CO2 at 37°C, the cells were then continuously cultured in fresh medium for 48 h for use in subsequent experiments. The lentiviral vector was used to construct the TGFβR2 overexpression vector: TGFβR2 cDNA with restriction enzyme sites of KpnI and XhoI, as well as a corresponding control sequence (designed by BLOCK-iT™ RNAi Deshgner website) was synthesized, and then ligated into the lentiviral expression vector pLVX-IRES-neo (Clontech Laboratories, Inc.) by T4 ligase. Following 24 h of culture at 37°C, the expression vector was extracted and sequenced. Finally, the packaged vector and viral particle were used to infect EC cells (2×104 cells) cultured in 5 µg/ml Polybrene cultured in a 12-well plate with a multiplicity of infection (MOI) of 50. At 24 h following transfection, cells were harvested for analysis.
Mir 93 5p mimic
Manufactured by RiboBio
Sourced in China
The MiR-93-5p mimic is a synthetic double-stranded RNA molecule that mimics the mature sequence of the microRNA miR-93-5p. MicroRNAs are small, non-coding RNA molecules that play a role in regulating gene expression. The MiR-93-5p mimic can be used for research purposes to study the biological functions of this specific microRNA.
Automatically generated - may contain errors
Lab products found in correlation
Mir 93 5p inhibitor, by RiboBio (4 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Plvx ires neo, by Takara Bio (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Control mirna, by RiboBio (1 mentions)
Mimic nc, by RiboBio (1 mentions)
Inhibitor nc, by RiboBio (1 mentions)
6 protocols using mir 93 5p mimic
1
Regulation of EC Cells by miR-93-5p and TGFβR2
2
XIST-mediated Regulation of miR-93-5p in Cancer
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For cell experiments, the plasmid based siRNA targeting for XIST and XIST overexpression vector were purchased from Shanghai Genepharma Co., Ltd. siRNA-XIST: 5′-AATGGAACGGGCTGAGTTTTAG-3′. MiR-93-5p mimics, miR-93-5p inhibitor and miRNA control were purchased from RiboBio Co., Ltd., Guangzhou, China. Briefly, 100 nM of MiR-93-5p mimics or miR-93-5p inhibitor or 1000 ng plasmid were transfected with each 6-well plate for 48 h using Lipofectamine 2000 (Invitrogen, Waltham, MA, USA). For animal experiments, cells were stably transfected with XIST based on lentiviral, constructed by Hanyin Biotechnology Co., Ltd., Shanghai, China. After transfection, the DMEM was replaced with DMEM supplemented with puromycin (3 μg/ml) for excluding non-infected cells. Then, the selected cell were used in the subcutaneous xenograft.
3
Modulating METTL3, miR-93-5p, and FRMD6 in PCa Cells
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METTL3-targeting siRNAs, miR-93-5p mimics, and miR-93-5p inhibitors were obtained from RiboBio (China) and transfected into PCa cells using a transfection reagent, Lipofectamine 3000 (Invitrogen, USA), following the manual provided by the manufacturer. The lentiviral vector containing the FRMD6 sequence and negative control were constructed by RiboBio (China) to infect DU145 and PC3 cells and obtain stable FRMD6-overexpressing cell lines after screening with puromycin. All the information for the above sequences is listed in Table S4 .
4
Regulating lncRNA H19 and miR-93-5p in hBSMCs
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The expression of lncRNA H19 in hBSMCs was inhibited with transfection of siRNA against H19 (50 nM). Scrambled siRNA was used as the negative control (Qijing, Wuhan, China). SiRNAs were diluted in riboFECTTMCP reagents (Ribobio, Guangzhou, China). Sequences of H19 siRNA were as follows: sense 5’-CCAACAUCAAAGACACCAUTT-3’, antisense 5’-AUGGUGUCUUUGAUGUUGGTT-3’. The expression of miR-93-5p was upregulated by miR-93-5p mimic and downregulated by miR-93-5p inhibitor (Ribobio, Guangzhou, China) respectively. miR-93-5p mimic or inhibitor was transfected into hBSMCs with riboFECTTMCP reagents. For transfection, hBSMCs were inoculated into 24-well plates with a density of 1×104 cells/well and transfection was conducted when cells grew to 50~60% confluence. The transfection complex was added into DMEM/F12 containing 10% FBS with a final volume of 500 μL. Control cells were treated only with same volume of PBS. The medium containing transfection complex was removed after 6 h, and cells were cultured in DMEM/F12 in the presence of IL-13 for an additional 24 − 72 h. The silencing efficiency of siRNA was examined with qRT-PCR.
5
Modulating lncRNA NORAD in HUVECs
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HUVECs were gained from the American Typical Culture Conservatory (ATCC), which were cultured in DMEM maintained in a 37 °C, 5% CO2 incubator. 10% fetal bovine serum and 10% endothelial cell growth supplement were added to the medium. Small interfering (si) RNA (si-NORAD) sequence of lncRNA NORAD gene and its negative control (si-NC), and overexpression plasmid of pcDNA3.1-NORAD were provided by GenePharma Co, while miR-93-5p mimic, mimic-NC, miR-93-5p inhibitor and inhibitor-NC were obtained from the RIBOBIO Co. When cells grew to 80% confluence, the cell transfection was performed using Lipofectamine 3000 (Invitrogen, USA). After 5 h of culture, a new medium was replaced.
6
Gastric Cancer Cell Lines: Inhibitor and Mimetic Effects
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Four human GC cell lines SUN-216 (CBP60503, culture condition: RPMI-1640 + 10% FBS), BGC-823 (CBP60477, culture condition: RPMI 1640 (w/o Hepes) + 10% FBS), MKN74 (CBP60490, culture condition: RPMI-1640 + 10% FBS) and HGC-27 (CBP60480, culture condition: MEM + 1%NEAA + 10% FBS) and human gastric epithelial cells GES-1 (CBP60512) were purchased from Nanjing Cobioer Biotechnology Co., Ltd. (Nanjing, China). Cells were assigned into the following groups: inhibitor-negative control (NC) group (cells treated with inhibitor-NC vector, purchased from Guangzhou RiboBio Biotechnology Company, Guangzhou, China), miR-93-5p inhibitor group (cells treated with miR-93-5p inhibitor vector), Empty vector group (cells treated with pCDH vector), AHNAK group (cells treated with pCDH-AHNAK vector), miR-93-5p mimic + Empty vector (cells treated with miR-93-5p mimic + pCDH vector), miR-93-5p mimic + AHNAK (cells treated with miR-93-5p mimic + pCDH-AHNAK-vector), miR-93-5p mimic + DMSO group (cells treated with miR-93-5p mimic + Dimethylsulfoxide (DMSO) vector), and miR-93-5p mimic + DKK1 (cells treated with miR-93-5p mimic + Wnt signaling pathway inhibitor DKK1 vector).
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